Engineering of a red-light–activated human cAMP/cGMP-specific phosphodiesterase

Abstract

Significance Sensory photoreceptors not only enable organisms to derive spatial and temporal cues from incident light but also provide the basis for optogenetics, which denotes the manipulation by light of living systems with supreme spatial and temporal resolution. To expand the scope of optogenetics, we have engineered the light-activated phosphodiesterase LAPD, which degrades the ubiquitous second messengers cAMP and cGMP in a red-light–stimulated manner. Both cAMP and cGMP are key to the regulation of manifold physiological responses, and LAPD now augurs red-light control over these processes. As we demonstrate for two eukaryotic systems, LAPD does not require any additional exogenous factors, and thus permits the light perturbation of living cells in hitherto unrealized ways.

Three cyanobacteriochromes work together to form a light color-sensitive input system for c-di-GMP signaling of cell aggregation

“…Generally, proteins containing active GGDEF and EAL domains are dimeric and their dimerization interfaces often modulate their DGC and PDE activities (17,31,32). GAF domains including those of CBCRs often transduce the input signal via a rotary movement of a connecting α-helix toward a neighboring output domain (41,42). Further characterization of SesB and SesC should clarify their intramolecular signaling and any possible intermolecular signaling with interacting proteins, which then might be used to design chimeric sensor proteins.…”

Phytochromes: An Atomic Perspective on Photoactivation and Signaling

“…However, other signaling motifs are also employed, including diguanyl cyclase, diguanylate phosphodiesterase, methyl-accepting, phosphatase 2C, two-helix output sensor, and histidine kinase/methyl-accepting/phosphatase (HAMP) domains, demonstrating that the Phy-type PSM is amenable to actuating a wide variety of other signaling pathways ( Figure 1). In fact, it is possible to attach the PSM of Phys to non-native signal output motifs to invent novel light-responsive pathways (Gasser et al, 2014;Ryu et al, 2014). Phenotypic outputs from these microbial Phys include the regulation of pigmentation, photoprotection, redox sensing, phototaxis, sporulation, entrainment of circadian rhythms, and the chromatic acclimation of the photosynthetic machinery to the available light spectrum (Auldridge and Forest, 2011;Bussell and Kehoe, 2014).…”

Synthetic biology of cell signaling

“…The result was a red-light responsive rhodopsin derived from Halobacterium salinarum cruxhalorhodopsin chloride pump, Jaws, that could hyperpolarize cells causing inhibition in brain neurons at a depth of 3 mm (Chuong et al 2014). Gasser et al (2014) also developed a red-light activated phosphodiesterase with the function of performing cAMP and cGMP hydrolysis; however, due to the nature of the bacterial system from which it originates redlight (650-700 nm) would increase hydrolysis up to six fold while far-red light (700-750 nm) could decrease activity. Both systems exploit synthetic systems and depth of red light penetration for use in optogenetics.…”